Cultivo in vitro de raíces pilosas del arbusto Phyllanthus acuminatus (Phyllanthaceae) / In vitro culture of hairy roots of the shrub, Phyllanthus acuminatus (Phyllanthaceae)

Introducción: En condiciones naturales, las raíces del arbusto, Phyllanthus acuminatus, producen bajas concentraciones de metabolitos secundarios de interés medicinal. Esto abre una oportunidad para el cultivo in vitro, para aumentar la concentración de metabolitos. Objetivo: Determinar las condiciones óptimas de cultivo líquido para raíces pilosas de P. acuminatus. Métodos: Se utilizó la evaluación del crecimiento de la biomasa según porcentaje de inóculo inicial (0.50 y 0.10 %), porcentaje de nutrientes de los medios (100, 50 y 25 %) y tasa de agitación (90, 100 y 110 min-1) (N= 15 repeticiones). Resultados: Las mejores condiciones de cultivo líquido fueron: 0.10 % de inóculo inicial, nutrientes al 25 % y 90 min-1 para la tasa de agitación. Hay diferencias entre las raíces pilosas y las raíces no transformadas. Conclusiones: es factible producir raíces pilosas de P. acuminatus a gran escala, aplicando e implementando las condiciones evaluadas de porcentaje de inóculo, nutrientes en el medio y tasas de agitación utilizadas en este estudio.

Introduction: Under natural conditions, the roots of the shrub, Phyllanthus acuminatus, produce low concentrations of secondary metabolites of medicinal interest. This opens an opportunity for in vitro culture, to increase metabolite concentration. Objective: To determine the optimal liquid culture conditions for hairy roots of P. acuminatus. Methods: We used biomass growth evaluation according to initial inoculum percentage (0.50 and 0.10 %), percentage of medium nutrients (100, 50 and 25 %) and agitation rate (90, 100 and 110 min-1) (N=15 replications). Results: The best liquid culture conditions were: 0.10 % of initial inoculum, nutrients at 25 % and 90 min-1 for the agitation rate. There are differences among hairy roots and non-transformed roots. Conclusions: It is feasible to produce P. acuminatus hairy roots at a large scale, applying and implementing the evaluated conditions of inoculum percentage, nutrients in the medium and agitation rates.

Influence of root-knot nematode, Meloidogyne incognita (Kofoid & White) Chitwood infection on different plant growth parameters in Mungbean, Vigna radiata (L.) Wilczek

Root-knot nematode, Meloidogyne incognita (Kofoid & White) Chitwood, is a major threat to mungbean cultivation. The pest causes a significant reduction in plant growth parameters that ultimately results in loss of grain yield. The present study was carried out under glass house condition to study the effect of different inoculum load of root-knot nematode M. incognita on plant growth, nodulation and nematode development and nutrients status of Mungbean. The results revealed a progressive decline in plant growth parameters viz., fresh and dry shoot weight and shoot length with respect to increase in inoculum level. However, fresh and dry root weight showed the opposite trend. The fresh and dry shoot weight was decreased by 44% and 66%, respectively at 4 J2s/g soil. The chlorophyll content in the leaves also decreased with the increase of inoculum level from 100-6000 J2s/pot. Nutrients contents of the plant viz. N, P, K, Ca and Mg were significantly reduced in shoots while in roots these was increased with an increase of inoculum levels. Nodulation was affected by 80% at the highest inoculum level i.e. 6000 J2s/pot. Also leghaemoglobin, bacteroid content and nitrogenase activity was reduced progressively with increased levels of nematode inoculum. Thus, the root-knot nematode, M. incognita interferes with the process of symbiotic nitrogen fixation between mungbean host and rhizobium and that can affect the quality of produce.

Potential of microbial inoculum from buffalo feces in activating lignite coal bed methane

Aim: This research aimed to study the ability of microbial consortium from buffalo feces to produce coal bed methane (CBM) in lignite coal through in-vitro technique.Methodology: This study was carried out in 2 stages: In first stage, microbes activated using in-vitro technique, and at second stage volatile fatty acids (VFA) and coal bed methane production were estimated. The nutrition source for inoculum activation was of three types: 100% concentrate: 70% concentrate: 30% grass and 30% concentrate: 70% grass. Biogas digester in laboratory-scale utilized 100 ml serum bottle filled with 70 ml, 98-5 media and 7 g coal. Microbial inoculum was inoculated on digester using a 10 ml syringe and incubated at 39oC. Parameters observed to measure the quality of inoculum were total number of anaerobic microbes, kinetics, and biogas production during fermentation. Complete Randomized Design with two factorial consisted of incubation period as factor A and inoculum dosages as factor B. Results: The activation process of inoculum from buffalo feces was required to produce coal bed methane in anaerobic digestion. During activation process, the microbes from buffalo feces in a mixture of 70% concentrate and 30% grass at 48 hr observation could produce total gas approximately 22.5 ml at 48 hr observation. Addition of activated buffalo feces anaerobic inoculum on the anaerobic digestion as much as 6% produced the highest number of anaerobic bacteria, and VFA accounted for 31 x 1010 CFU ml-1 and 171.7 mM, respectively. Meanwhile, the highest methane production reached 128.61 ml on adding 6% inoculum

Pre-incubation of ruminal inocula to assess in vitro gas production and digestibility / Pré-incubação de inóculo ruminal para avaliação da produção de gases e digestibilidade in vitro

ABSTRACT: In vitro gas production techniques represent a valuable tool to describe the kinetics of ruminal degradation of food. However, the ruminal liquor used as a microbial inoculum has been a great source of variation and error. A standardization of this factor should contribute to assure the independence of food fermentation parameters from those of the inocula. In this research it was hypothesized that a controlled pre-incubation treatment of ruminal liquor could contribute to stabilize and homogenize the undigested residues of blanks and as a consequence, of the production of residual cumulative gas production (CGP). A pre-incubation (i.e. previous real incubation) of rumen inocula was developed with a simple substrate similar to the diet offered to donors at 1% w/v for 0, 1, 2 and 4 h (Control, Prei-1, Prei-2 and Prei-4 treatments respectively). Once the pre-incubation hours were completed, they were incubated with contrasting substrates and without substrate (i.e. blanks) in order to evaluate the CGP, in vitro digestibility of the DM and fermentation products. Although, the fermentative activity of the pre-incubated inoculums worked satisfactorily in the in vitro system, contrary to what was speculated, residues of the pre-incubation increased the variability and heterogeneity of variances among blanks. Consequently, it was concluded that the pre-incubations did not work to generate more homogeneous and less variable ruminal liquor for the in vitro gas production system.

RESUMO: Técnicas de produção de gás in vitro representam uma ferramenta valiosa para descrever a cinética de degradação ruminal dos alimentos. No entanto, o líquido ruminal utilizado como inóculo microbiano tem sido uma grande fonte de variação e erro. A padronização deste fator deve contribuir para garantir a independência dos parâmetros de fermentação dos alimentos a partir dos inóculos. Neste trabalho, hipotetizou-se que um tratamento controlado de pré-incubação do líquido ruminal poderia contribuir para estabilizar e homogeneizar os resíduos não digeridos dos brancos e, como conseqüência, da produção de produção cumulativa de gás residual (CGP). Uma pré-incubação (ou seja, incubação real prévia) dos inóculos do rúmen foi desenvolvida com um substrato simples semelhante à dieta oferecida aos doadores a 1% p/v por 0, 1, 2 e 4 h (Controle, Prei-1, Pré- 2 e Prei-4 tratamentos respectivamente). Uma vez completadas as horas de pré-incubação, elas foram incubadas com substratos contrastantes e sem substrato (ou seja, brancos) para avaliar o CGP, a digestibilidade in vitro da MS e os produtos de fermentação. Embora a atividade fermentativa dos inóculos pré-incubados tenha funcionado satisfatoriamente no sistema in vitro, ao contrário do que foi especulado, os resíduos da pré-incubação aumentaram a variabilidade e heterogeneidade das variâncias entre os brancos. Consequentemente, concluiu-se que as pré-incubações não funcionaram para gerar um líquido ruminal mais homogêneo e menos variável para o sistema de produção de gás in vitro.

Isolation of phytoplasmas associated to frogskin disease in cassava / Aislamiento de fitoplasmas asociados a cuero de sapo en yuca

ABSTRACT Frogskin is the most limiting disease of cassava crops in Colombia, causing losses in production up to 90%. Since this disease was associatated with 16SrIII phytoplasma presence, a study was carried out to isolate this phytoplasma using liquid and solid culture media. Root, petiol, stem, leaf and cutting tissues of cassava affected by frogsking were employed as source materials. Molecular and microscopy techniques were applied to verify the phytoplasma growth and to discard other microorganism´s presence. The results showed that the media consistently allow phytoplasma growth, and colonies in solid medium were obtained. PCR, qPCR and sequencing tests confirmed the presence of 16SrIII group phytoplasmas in both liquid and solid culture media. Additionally, the isolation of a pigeon pea witches' broom phytoplasma strain (group 16SrIX) was obtained from stems, petioles and flowers of symptomatic Catharanthus roseus confirming the effectiveness of the medium in the phytoplasma isolation and culture. This is the first isolation of field-collected phytoplasma strains in groups 16SrIII and 16SrIX in America that confirm and corroborate the previous results in phytoplasma cultivation achieved on micropropagated and field-collected phytoplasma infected samples.

RESUMEN En Colombia, el ''cuero de sapo'' es la enfermedad más limitante del cultivo de yuca, que ocasiona pérdidas en producción de raíces hasta del 90%. La presente investigación tuvo como objetivo, el aislamiento in vitro del fitoplasma asociado a cuero de sapo. Para ello, se emplearon medios de cultivo líquido y sólido, usando tejidos de raíces, peciolos, tallos, hojas y semillas de yuca, afectada por la enfermedad. Pruebas de PCR, qPCR, secuenciación, microscopia de luz y microscopia electrónica de transmisión fueron aplicadas, para verificar el crecimiento de fitoplasmas y descartar la presencia de otros microrganismos. Los resultados muestran que los medios permiten, consistentemente, el crecimiento de fitoplasmas, obteniendo colonias en medio sólido a partir de medio líquido. Las pruebas de PCR, qPCR y secuenciación confirmaron presencia de Cassava frogskin phytoplasma del grupo 16SrIII, en los dos medios de cultivo. Además, a partir de las colonias, se lograron fotografías de células con morfología y tamaño similares a las fitoplasmáticas. Es la primera vez, en el mundo, que se consolida información suficiente del aislamiento de fitoplasmas en medio artificial. Adicionalmente, se logró el aislamiento de Pigeon pea witches´ broom phytoplasma del grupo IX, a partir de tallos, peciolos y flores de vinca (Catharanthus roseus), con síntomas asociados a fitoplasmas. Este proceso permitió corroborar la efectividad del medio y la morfología de las células fitoplasmáticas, bajo microscopia electrónica.

Efficacy of alginate- and clay-encapsulated microorganisms on the growth of Araçá-Boi seedlings (Eugenia stipitata)

The present work aimed to evaluate the effects of encapsulated microorganisms on seedlings of Eugenia stipitata, popularly known as araçá-boi, to evaluate the interaction between the inoculum and encapsulating agents such as clay and alginate. The experiment was carried out in a completely randomized design using a 3×2 factorial scheme. The treatments were control, inoculum, clay without microbial inoculum, clay with microbial inoculum, alginate without microbial inoculum, and alginate with microbial inoculum. The seedlings were grown under nursery conditions over a period of 3 months. No treatment increased the height, stem diameter, shoot dry matter or root dry matter of the araçá-boi seedlings. The use of alginate increased the ammonium content compared to the clay and control treatments. Alginate and clay increased the nitrate content in relation to the control. Alginate increased the total number of bacteria in relation to the clay and control treatments. The application of inoculum combined with alginate increased the nitrate content only in relation to the clay and control treatments. Although the application of inoculum promoted an increase in the nitrate content compared to the uninoculated treatments, there was no effect for the other parameters analyzed. The results suggest that clay and alginate encapsulating agents with the presence or absence of microorganisms may improve some soil parameters.

Inactivation of Alicyclobacillus acidoterrestris ATCC 49025 spores in apple juice by pulsed light: Influence of initial contamination and required reduction levels / Inactivación de esporas de Alicyclobacillus acidoterrestris ATCC 49025 por luz pulsada en jugo de manzana: Influencia de la contaminación inicial y de las reducciones requeridas

The purpose of this study was to analyze the response of different initial contamination levels of Alicydobadllus acidoterrestris ATCC 49025 spores in apple juice as affected by pulsed light treatment (PL, batch mode, xenon lamp, 3pulses/s, 0-71.6 J/cm²). Biphasic and Weibull frequency distribution models were used to characterize the relationship between inoculum size and treatment time with the reductions achieved after PL exposure. Additionally, a second order polynomial model was computed to relate required PL processing time to inoculum size and requested log reductions. PL treatment caused up to 3.0-3.5 log reductions, depending on the initial inoculum size. Inactivation curves corresponding to PL-treated samples were adequately characterized by both Weibull and biphasic models (R²d j 94-96%), and revealed that lower initial inoculum sizes were associated with higher inactivation rates. According to the polynomial model, the predicted time for PL treatment increased exponentially with inoculum size.

El objetivo del presente trabajo fue evaluar la influencia de la concentración de esporas de Alicyclobacillus acidoterrestris ATCC 49025 en la respuesta de inactivación por acción de la luz pulsada (modo estanco, lámpara de xenón, 3 pulsos/s, 0-71,6 J/cm²) en jugo de manzana comercial. Para caracterizar la relación existente entre la concentración de esporas y el tiempo de tratamiento con las reducciones logarítmicas alcanzadas luego de la exposición a la luz pulsada (LP), se aplicaron 2 modelos: el de Weibull y el bifásico. Adicionalmente, se estimó la relación entre el tiempo de tratamiento con LP y la concentración inicial de inoculo en el jugo con las reducciones logarítmicas logradas mediante regresión múltiple y la metodología de superficie de respuesta (MSR). La inactivación por LP provocó entre 3 y 3,5 reducciones logarítmicas, según la concentración inicial de esporas. Las curvas de inactivación fueron adecuadamente caracterizadas por los modelos matemáticos propuestos (Restado = 94-96%). El análisis por MSR permitió predecir un aumento exponencial del tiempo de tratamiento requerido conforme se incrementa el nivel de contaminación inicial.

Yield and rheological properties of exopolysaccharide from a local isolate: Xanthomonas axonopodis pv. vesicatoria

Background: The aim of the present study was to evaluate gum productivity of a local strain, Xanthomonas axonopodis pv. vesicatoria, isolated from pepper plant, and its rheological behavior for the first time compared to the standard strain, Xanthomonas campestris DSM 19000 (NRRL B-1459). The influence of operational conditions (agitation rate and inoculum volume) on gum production and rheological properties of gums from the Xanthomonas strains were investigated. Results: The isolated strain of Xanthomonas showed similar xanthan yield compared to the standard strain. Furthermore, this study clearly confirmed that gum yield depended on bacterial strain, agitation rate, and inoculum size. The most suitable conditions for the gum production in an orbital shaker in terms of agitation rate and inoculum size were 180 rpm and 5%, respectively, resulting in an average production of 10.96 and 11.19 g/L for X. axonopodis pv.vesicatoria and X. campestris DSM 19000, respectively. Regarding the rheological properties, Ostwald-de-Waele and power law models were used to describe flow and oscillatory behavior of the gum solutions, respectively. Consistency of the novel gum solution remarkably was much higher than the commercial xanthan gum solution. Flow and oscillatory behavior and their temperature ramps showed that weak gel-like structure could be obtained with less gum concentrations when the novel gum was used. Conclusion: Therefore, yield and technological properties of the aqueous solutions of the exopolysaccharide synthesized by X. axonopodis pv. vesicatoria were observed to be more suitable for industrial production.

Influence of manure concentration as inoculum in anaerobic digestion of vinasse / Influência da concentração de esterco como inóculo na digestão anaeróbia da vinhaça

Vinasse, main residue of the sugarcane industry, has high pollutant content, being subjected to the use in biogas production due to the high content of organic matter non-toxic to microbial action. For a consolidated process, it is necessary to study parameters that influence the process, in which the amount of inoculum is one of the major factors in the biological process of biogas production. This study investigated the influence of the amount of manure as inoculum (0.5 to 5.5%) during the biodigestion process, evaluating variables such as chemical oxygen demand (COD), pH, biogas production, methane concentration, total solids and total phosphorus and nitrogen contents, as well as microbiological analysis in the sludge remaining in the digester. Biodigestion occurred normally, with hydraulic retention time (HRT) of 20 days, with an acidogenic phase, subsequent stabilization of pH and biogas production. The vinasse had COD and total solids reduced during biodigestion by around 67 and 40%, respectively. Biogas production was increased after the fifth day. Among the three studied conditions, there was no significant increase in efficiency of inoculum use and it can be used the lowest amount, 0.5 % (m v-1).

A vinhaça, principal resíduo da indústria sucroalcooleira, possui alto teor poluente, sendo passível seu uso na produção de biogás devido ao grande teor de matéria orgânica não tóxica à ação microbiana. Para que haja um processo consolidado, é necessário estudar parâmetros que influem em seu processo, sendo a quantidade de inóculo um dos principais fatores no processo biológico de produção de biogás. Este estudo verificou a influência da quantidade de esterco bovino como inóculo (de 0,5 a 5,5%) durante o processo de biodigestão, avaliando variáveis como demanda química de oxigênio (DQO), pH, produção de biogás, concentração de metano, sólidos totais e teores de fósforo total e de nitrogênio total, além de análises microbiológicas no lodo restante no biodigestor. A biodigestão aconteceu normalmente, com um tempo de retenção hidráulica (TRH) de 20 dias, verificando-se a fase acidogênica, posterior estabilização do pH e produção de biogás. A vinhaça teve seus parâmetros de DQO e de sólidos totais diminuídos durante a biodigestão, em torno de 67 e 40%, respectivamente. Houve crescente produção de biogás após o quinto dia. Percebeu-se que, dentre as três condições estudadas, não houve significativa elevação da eficiência de uso de inóculo, podendo ser usada a menor quantidade, 0,5% (m v-1).

Production of native arbuscular mycorrhizal fungi inoculum under different environmental conditions

Abstract In order to obtain an arbuscular mycorrhizal fungi (AMF) native inoculum from Sierra de Moa and determine the most appropriate conditions for its big scale production, four light and temperature combinations were tested in three plant species (Calophyllum antillanum, Talipariti elatum and Paspalum notatum). Growth and development parameters, as well as the mycorrhizal functioning of the seedlings were evaluated. The natural light treatment under high temperatures (L-H) was the most suitable for the growth and development of the three plant species, showing the highest total biomass values, mainly of root, and a positive root-shoot ratio balance. This treatment also promoted higher values of root mycorrhizal colonization, external mycelium and AMF spore density. A total of 38 AMF species were identified among the plants and environmental conditions tested. Archaeospora sp.1, Glomus sp.5, Glomus brohultii and G. glomerulatum were observed in all the treatments. The L-H condition can be recommended for native inoculum production, as it promotes a better expression of the AM symbiosis and an elevated production of mycorrhizal propagules.

Microbial Contamination and Evaluation of Inoculum Volume in Umbilical Cord Blood Culture / 대한임상미생물학회지

BACKGROUND: Microbial screening tests of umbilical cord blood (UCB) are essential for stem cell transplantation. We analyzed the microbial contamination rate and distribution of isolated microorganisms over 10 years of samples from the MEDIPOST Cord Blood Bank. In addition, we studied the influence of inoculum volume microorganism culture and compared the yield and speed of microorganism detection. METHODS: Microbial screening tests were performed using a manual method, which includes using an inoculum of 2 mL of plasma, a byproduct of UCB processing from pediatric culture bottles. When positive blood culture was detected, each set was once again inoculated with 2 mL and 4 mL of plasma. RESULTS: From 2004 to 2013, a total of 133,610 UCB units were screened, of which 1,311 (0.9%) tested positive for contamination. The most frequently identified microorganism was Escherichia coli (34.6%), followed by Bacillus spp. (12.8%), Enterococcus faecalis (5.3%) and Klebsiella pneumoniae (4.4%). The total yield rate increased by 0.2% over this time period, although the yield rate of Bacillus spp. increased by 8.3%. CONCLUSION: The results of this study could be used in many ways with both domestic and international data regarding cord blood contamination. Also, other microbiology laboratories using culture conditions similar to ours could refer this study when preparing guidelines. Finally, by detecting low levels of bacteria, we have contributed to cord blood safety.

PATHOGENICITY OF Blastocystis sp. TO THE GASTROINTESTINAL TRACT OF MICE: RELATIONSHIP BETWEEN INOCULUM SIZE AND PERIOD OF INFECTION / Histopatogenicidade de Blastocystis sp. para o trato gastrointestinal de camundongos: relação com inóculo e tempo de infecção

The pathogenic potential of Blastocystis sp. in experimental models requires further investigation. In this work, the pathogenicity of this parasite in the gastrointestinal tract of male Swiss mice was evaluated according to the inoculum size and period of infection. Animals were infected intragastrically, with 100, 500, 1,000, 5,000 and 10,000 Blastocystis sp. vacuolar forms obtained from a mixture of eight human isolates cultured axenically in Jones' medium. After seven, 14, 21, 28 and 60 days of infection, the animals were sacrificed and fragments of the small intestine (duodenum), large intestine, and cecum were subjected to histopathological analysis. Blastocystis sp. triggered an inflammatory response in the different tissues analyzed, with a predominance of mononuclear cells. The parasite was found in the muscular layer of the cecum, showing its invasive character. Larger inocula triggered inflammatory processes earlier (seven days) than smaller ones (from 21 days). We conclude that, in the proposed model, the pathogenicity of Blastocystis sp. isolates that were studied is related to inoculum size and period of infection.

Pouco é sabido sobre o potencial patogênico de Blastocystis sp. em modelos experimentais. Neste trabalho a patogenicidade desse parasito para o trato gastrointestinal de camundongos Swiss machos foi avaliada de acordo com o inóculo e tempo de infecção. Os animais foram infectados, via intragástrica, com 100, 500, 1.000, 5.000 e 10.000 formas vacuolares de Blastocystis sp. obtidos a partir de uma mistura de oito isolados humanos cultivados axenicamente em meio Jones. Após 7, 14, 21, 28 e 60 dias de infecção os animais foram sacrificados e fragmentos do intestino delgado (duodeno), grosso e ceco foram retirados para análise histopatológica. Blastocystis sp. desencadeou resposta inflamatória nos diferentes tecidos analisados, com predominância de infiltrado mononuclear. No ceco o parasito foi encontrado na túnica muscular mostrando seu caráter invasivo. Inóculos maiores desencadearam processos inflamatórios mais precocemente (7 dias) e inóculos menores mais tardiamente (a partir de 21 dias). Conclui-se que no modelo proposto a patogenicidade dos isolados de Blastocystis sp. estudados tem relação com o inóculo e tempo de infecção.

A method for early detection of antibiotic resistance in positive blood cultures: Experience from an oncology centre in eastern India.

Purpose: For antibiotic susceptibility results, conventional culture and sensitivity methods takes 48 hours after a blood culture is fl agged positive by automated systems. Early initiation of targeted antibiotic therapy is essential for effective management of sepsis to reduce morbidity, mortality, cost of treatment and prevent antibiotic resistance. Objective of this study was to evaluate Direct Sensitivity Test (DST) as a potential tool to get reliable antibiotic susceptibility results 24 hours earlier. Materials and Methods: Blood cultures fl agged positive between May 2011 to December 2012 by BacT/ALERT were Gram stained. All uni-microbial gram-negative blood cultures were simultaneously cultured and processed for DST from broth using disk diffusion method using British Society of Antimicrobial Chemotherapy (BSAC) guidelines. DST results available next day were compared with conventional antibiotic susceptibility test (AST) performed by Vitek-2 on isolated colonies. Results of DST (test method) and AST (reference method) were compared for agreements or errors. Results: Of the 840 antibiotic gram-negative organism combinations tested, Categorical and essential agreements were 83.7% and 96.2% respectively. Minor, major and very major errors were 12.5%, 3.33% and 0.47%, respectively. Conclusions: DST using disk diffusion from positive blood culture broths helps to initiate early targeted antibiotic therapy. There is high concordance between DST and AST.

Efecto de la carga orgánica de la gallinaza de jaula en el potencial de biometanización / Effect of chicken manure organic load on biomethane potential

El propósito de este estudio fue evaluar el efecto de la carga orgánica expresada en función de la relación inóculo/sustrato (RIS) sobre el potencial de biometanización de la gallinaza de jaula usando como inóculo lodo estiércol bovino. Se llevaron a cabo ensayos de biodegradación anaerobia a temperatura mesofílica de 39 °C. Para cada una de las cargas orgánicas evaluadas (16.6, 11.0, 8.3, 6.6 y 5.5 g SV/L) se cuantificaron las variables: ácidos grasos volátiles totales, alcalinidad, concentración amonio y volumen acumulado de metano. El mayor potencial de biometanización (0.58 m³ CH4/kg SV) se alcanzó cuando la biodegradación anaerobia se llevó a cabo con una RIS de 1.0. Los resultados obtenidos demuestran que la gallinaza es un sustrato potencial para ser degradado por digestión anaerobia y el rendimiento del proceso es directamente proporcional a la concentración de sustrato. Este estudio también confirma que la RIS permite diluir la concentración de compuestos inhibitorios como el amonio en el caso de la gallinaza de jaula.

The aim of this study was to evaluate the effect of inoculum to substrate radio (ISR) on biomethane potential of chicken manure using cattle slurry as inoculum. Biomethane potential assays were carried out at 39 °C mesophilic temperature. Total fatty acids, total alkalinity, ammonium concentration and accumulative methane volume were measured to evaluate organic load (16.6, 11.0, 8.3, 6.6 y 5.5 g VS/L). The highest biomethane potential (0.58 m³ CH4/kg SV) was reached when anaerobic biodegradation was carried out to ISR of 1.0. The results demonstrated that chicken manure is a potential substrate to be degraded by anaerobic digestion and process performance is directly proportional to substrate concentration. This study also confirms that ISR allow dilution of inhibitory components as ammonium by the case of chicken manure.

Inoculum Sizes of Locally Isolated Phototrophic Bacterium on the Utilization of Palm Oil Mill Effluent.

Aims: To produce Rhodobacter sphaeroides strain UMSPSB3 biomass with the reduction of chemical oxygen demand (COD) from palm oil mill effluent. Study Design: Locally isolated phototrophic bacterium with different inoculum levels were used in Palm Oil Mill effluent (POME). Collected POME was characterized before used as substrate. Inoculum of bacterium was grown in synthetic media and 48 hours inoculum was used to utilize the substrate. Place and Duration of Study: Biotechnological laboratory, Borneo Marine Research Institute, University Malaysia Sabah, Kota Kinabalu, Sabah, Malaysia, between February 2014 to April 2014. Methodology: Growth characteristics of bacterium Rhodobacter sphaeroides strain UMSPSB3 was monitored at different light intensities. Later phototrophic bacterium Rhodobacter sphaeroides strain UMSPSB3 was grown in settled non-sterilized Palm Oil Mill effluent (POME). The growth characteristics of bacterium in term of dry cell weight and total carotenoids production, and reduction of COD were compared using 10%, 20% and 30% (v/v) levels of inoculum developed in synthetic 112 media. Results: The optimum light intensity for the growth of Rhodobacter sphaeroides strain UMSPSB3 was 2.5 klux. The highest bacterial biomass (Xmax) of 6.5 g/L (dry weight) and 72% reduction of COD were obtained after 96-h culture with 20% (v/v) inoculum level. The reduction of COD (%) and cell yield (Yx/y, g cell/g COD) in POME were 82% and 0.98 respectively, after 96-h culture with 30% (v/v) inoculum. Production of carotenoids was comparatively low in bacterium using POME as substrate. Inoculum levels of 20-30% (v/v) developed in synthetic 112 media supported the growth of phototrophic bacterium in settled POME, but higher level of inoculum was required for faster removal COD from effluent. A 10% (v/v) level of inoculum in POME did not support the isolate to grow. Conclusion: Production of bacterial biomass with bioremediation of effluent could be achieved using POME as substrate with locally isolated Rhodobacter sphaeroides strain.

The inoculum effect of fosfomycin in tests with Enterococcus / 中华传染病杂志

Objective To investigate the inoculum effect of fosfomycin on enterococci,in comparison with penicillin,rifampin,vancomycin and teicoplanin.Methods A total of 91 strains of enterococci were obtained from the patients hospitalized in The First Affiliated Hospital of China Medical University from January 2010 to January 2012.There was no repeated strain.The 91 strains consisted of 44 Enterococcus faecalis and 47 Enterococcus faecium strains.Agar dilution method was used to measure the minimum inhibitory concentrations (MIC) of the five antibiotics against the 91 enterococci strains under standard low inoculation concentration (1 × 105 colony-forming units [cfu]/mL) and high inoculation concentration (1 × 107 cfu/mL).Results For all tested strains,MIC of fosfomycin fluctuated between 16-64 tμg/mL in high and low inoculated concentrations.Among the tested strains,both Enterococcus faecalis and Enterococcus faecium had the highest drug-sensitive rate (100.0％) and the lowest drugresistance rate (0) to fosfomycin.With the increase of the inoculation concentration,MIC of the five antibiotics increased less than 8 times,which indicated that there was no inoculation effect.Conclusion Fosfomycin shows high drug-sensitivity against enterococci without inoculum effect in vitro,and it is recommended for the treatment of enterococci-infected patients.

Optimization of the production of mycorrhizal inoculum on substrate with organic fertilizer

The system for production of inoculum of arbuscular mycorrhizal fungi (AMF) using sand and vermiculite irrigated with nutrient solution is promising. However, organic amendments added to the substrate can stimulate sporulation of AMF and replace the nutrient solution. The aim of this study was to maximize the production of AMF (Acaulospora longula, Claroideoglomus etunicatum, Dentiscutata heterogama and Gigaspora albida) using selected organic substrates (vermicompost, coir dust and Tropstrato) together with sand and vermiculite. The production of spores varied among the tested AMF and according to the organic source added to the substrate. The vermicompost promoted higher sporulation of A. longula in relation to the other AMF and substrates. The Tropstrato® inhibited the sporulation of D. heterogama while the reproduction of C. etunicatum was not affected by the organic compounds. The inoculum of A. longula also showed a high number of infective propagules and promoted biomass accumulation in maize plants. The system of inoculum production using sand and vermiculite + 10% vermicompost favors the production of infective inoculum of A. longula with the fungus benefiting growth of corn plants.

DETERMINACIÓN IN VITRO DE LA ACCIÓN PROBIÓTICA DE Lactobacillus plantarum SOBRE Yersinia pseudotuberculosis AISLADA DE Cavia porcellus / IN VITRO DETERMINATION OF THE PROBIOTIC Lactobacillus plantarum ACTION ON Yersinia pseudotuberculosis ISOLATED FROM Cavia porcellus

Se evaluó el efecto in vitro de Lactobacillus plantarum sobre Yersinia pseudotuberculosis aislada de cuyes (Cavia porcellus) e identificada bioquímica y molecularmente. Se determinó la viabilidad de L. plantarum a diferentes pH (3.25, 4.5, 7.6 y 8.2), sales biliares bovinas (3, 4 y 5%), bilis bovina (0.5, 1 y 2%) y temperatura (38 y 45°C). Se realizó antibiograma a ambas bacterias con Cefalotina, Cefepima, Ciprofloxacina, Dicloxacilina, Enrofloxacina, Gentamicina, Penicilina y Trimetropim-Sulfametoxasol. Se determinaron los péptidos de L. plantarum mediante HPLC y la cinética de crecimiento en dos sustratos (MRS y Pro), midiéndose: biomasa, pH, consumo de azúcar total y ácido láctico. Se observó un mejor crecimiento bajo pH 3.25, sales biliares 5%, bilis bovina 2% y temperatura de 45°C con valores de 3.0·10(12), 2.3·10(12), 8.6·10(11) y 3.0·10(12) UFC/ml, respectivamente. L. plantarum y el sobrenadante inhibieron a Y. pseudotuberculosis, con halos de 3 y 5 mm. Se observó resistencia de L. plantarum y Y. pseudotuberculosis a Penicilina y Dicloxacilina, respectivamente, con sensibilidad de ambas cepas a los demás antibióticos. Se evidenciaron los péptidos VAL-TIR-VAL y TIR-GLI-GLI-FA-MET en el sobrenadante de L. plantarum. Durante la fase logarítmica para los medios MRS y Pro se alcanzaron valores de 6.75·10(12) y 1.20·10(12) UFC/ml (biomasa), 6.96 y 5.49 (pH), y 0.65 y 0.76%, (ácido láctico), respectivamente. La comparación de los medios mediante un diseño de bloques al azar (P > 0.05), mostró similar comportamiento. L. plantarum demostró su potencial probiótico en condiciones in vitro.

The in vitro effect of Lactobacillus plantarum on Yersinia pseudotuberculosis isolated from guinea pig identified and evaluated biochemical and molecular. L. plantarum viability was determined at different pH (3.25, 4.5, 7.6 y 8.2), bovine bile salts (3, 4 y 5%), bovine bile (0.5, 1 y 2%) and temperature (38 y 45°C). Both bacteria susceptibility was performed with Cephalothin, Cefepime, Ciprofloxacin, Dicloxacillin, Enrofloxacin, Gentamicin, Trimethoprim-Sulfamethoxazole and Penicillin. L. plantarum peptides were determined by HPLC and growth kinetics on two substrates (MRS and Pro), measuring biomass, pH, total sugar consumption and lactic acid. Better growth was observed at 3.25 pH, 5% bile salts, 2% bovine bile and temperature 45 °C. with values 3,0·10(12), 2.3·10(12), 8.6·10(11) y 3.0·10(12) UFC/ml, respectively. L. plantarum and the supernatant inhibited Y. pseudotuberculosis, with halos of 3 to 5 mm. L. plantarum and resistance to penicillin and Y. pseudotuberculosis Dicloxacillin was observed, respectively, with sensitivity of both strains to other antibiotics. Peptides were evidenced VAL-TIR-VAL y TIR-GLI-GLI-FA-MET in the supernatant of L. plantarum. During the logarithmic phase for MRS and Pro 1012 values were reached 6.75·10(12) y 1.20·10(12) CFU / ml (biomass), 6.96 and 5.49 (pH) and 0.65 and 0.76 % (lactic acid) respectively. The comparison of means using a randomized block design (P > 0.05) showed similar behavior. L. plantarum demonstrated their probiotic potential in vitro conditions.

Physiological Characterization of Fungal Inoculum for Biotechnological Remediation of Soils

The aim of this work was to study the bioremediating potential of Lentinus crinitus CCIBt2611 according to the physiological condition of the inoculum. Inoculum was prepared using sugarcane ground husk (C:N 90), at several physiological ages and applied in soil contaminated with pentachlorophenol. The inoculum's potential was assessed by evaluating the mycelium's vigor at soil's colonization, determination of peroxidase and phenoloxidase activities, in vitro degradation of Remazol Brilliant Blue R and in vivo degradation of pentachlorophenol. The results showed that the assessed parameters were relevant to identify the quality of the inoculum. For L. crinitus, 10 day old inoculum showed good soil-colonization speed with significant enzymatic activities, indicating the role of Manganese-dependent peroxidase and laccase in degradation, and efficient degradation of pentachlorophenol.

Producción a nivel de laboratorio de Clostridium septicum IRP15 para la formulación de una vacuna veterinaria / Laboratory production of Clostridium septicum IRP15 for the formulation of a veterinary vaccine

Objetivo: establecer las condiciones de producción a nivel de laboratorio de la alfa toxina de Clostridium septicum IRP15 para la formulación de una vacuna veterinaria y la optimización del proceso de producción. Métodos: se caracterizó y estandarizó la edad apropiada del inóculo para los cultivos en un fermentador New Brunswick Scientific 7 L. Las condiciones de cultivo fueron: cepa C. septicum IRP15, medio de cultivo VBH, 5 L/vaso de 7 L, inóculo de 250 mL (5 por ciento), 37 ºC, 24 h, bajo agitaciones prueba de 0, 25 y 50 r.p.m. Se estableció el perfil cinético morfológico, de biomasa, consumo de sustrato y producción de toxina. Resultados: para las fermentaciones de 0 y 25 r.p.m. no se presentó fase de adaptación; el microorganismo creció de manera exponencial hasta las 4 y 6 h de fermentación, consumiendo simultáneamente la mayor cantidad de glucosa presente en el medio. A partir de estas horas y hasta las 24, se realizó la prueba de DL50 en ratones y se destaca que a 25 r.p.m. se obtuvo el mayor título de toxina (1/23). En las fermentaciones a 50 r.p.m. se observó que el microorganismo experimenta una fase de adaptación de 4 h aproximadamente; con un retardo en producción de biomasa, consumo de glucosa y producción de toxina, condición que no resulta óptima para la producción del antígeno. Conclusiones: la producción de toxina se presenta en la fase logarítmica y durante la fase estacionaria, asociándose así al crecimiento y al fenómeno de esporulación(AU)

Objective: to set the laboratory production conditions of Clostridium septicum IRP15 alpha toxin for the formulation of a veterinary vaccine and the optimization of the productive process. Methods: the appropriate inoculum age for the cultures was characterized and standardized in a 7L New Brunswick Scientific biorreactor. The conditions of culturing were C. septicum IRP15 strain, VBH medium at 5 L/7 L glass, 250 mL (5 percent) inoculum, 37 ºC, and 24 h under shaking conditions of 0, 25 y 50 r.p.m. The following kinetic parameters were monitored: morphological changes, biomass production, glucose consumption and toxin production. Results: for the shaking conditions at 0 and 25 r.p.m., C. septicum did not show an adaptation phase growth. The bacteria kept growing at the log phase up to 4-6 hours of fermentation respectively, thus consuming the highest amount of glucose from the medium. As from the growth phase hours till the 24 h of cultivation, the 50 percent lethal dose (LD50) in mice assay was conducted and at 25 r.p.m. condition, the best titre of toxin was reached (1/23). The cultures at 50 r.p.m. condition showed that the bacteria experienced adaptation phase for almost four hours, resulting in delayed biomass production, glucose consumption and toxin production. These results suggested that 50 r.p.m. is not useful for the antigen production. Conclusions : the toxin production occurred at the log phase and during the stationary phase, thus it is associated to growth and to sporulation(AU)